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Studies on the pathogenesis and the treatment of GM1-gangliosidosis mice

GM1神经节苷脂沉积症小鼠发病机制及治疗研究

基本信息

批准号：
10680784
负责人：
MATSUDA Junichiro
金额：
$ 2.05万
依托单位：
National Institute of Infectious Diseases
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

GM1-gangliosidosis is a progressive neurological disease caused by a deficiency of lysosomal β-galactosidase(β-Gal) and is classified into three clinical forms : infantile, juvenile and adult. We recently generated β-Gal knockout(KO) mice as an authentic model of GM1-gangliosidosis. In this study, we conducted biochemical analysis of the model mice and demonstrated that the accumulation of gangliosides GM1 and asialoGM1 occurred as early as 1-2 weeks of age in their brains. We also generated the C57BL/6 congenic strain of KO mice and revealed the some differences in. storage materials between original crossbred and congenic strains. To evaluate the gene therapy using adeno-associated virus(AAV) vector for GM1-gangliosisosis mice, we injected AAV containing human β-Gal into the mouse brain and demonstrated the enzyme activities were detected only in a limited area for one month. We introduced the wild-type human β-Gal transgene into β-Gal KO mice in order to rescue their GM1-gangliosidosis phenotypes. We also introduced into β-Gal KO mice the mutant human β-Gal transgenes with R201C and I51T amino acid substitutions, which were common for juvenile and adult GM1-gangliosidoses, respectively. Three types of transgenes containing the β-actin promoter/human β-Gal cDNA (wild-type, R201C or I51T) were used. Transgenics were bred with β-Gal KO mice, and the resulting three types of KO/transgenic mice lacked mouse β-Gal gene but had human β-Gal (wild-type, R201C or I51T) transgene. As expected, the wild-type human β-Gal transgene could rescue the mouse GM1-gangliosidosis. KO/transgenic mice expressing mutant human β-Gals might be useful for the studies of pathogenesis and treatment of GM1-gangliosidosis with residual enzyme activities.

GM 1-神经节苷脂沉积症是一种进行性神经系统疾病，由溶酶体β-半乳糖苷酶（β-Gal）缺乏引起，分为三种临床形式：婴儿型、青少年型和成人型。我们最近产生了β-Gal敲除（KO）小鼠作为GM 1-神经节苷脂沉积症的真实模型。在这项研究中，我们对模型小鼠进行了生化分析，证明神经节苷脂GM 1和去唾液酸GM 1的积累早在1-2周龄时就发生在它们的大脑中。我们还建立了C57 BL/6同源株KO小鼠，并揭示了一些差异。原交系与同源系之间的贮藏材料。为了评价腺相关病毒（AAV）载体对GM 1-神经节细胞变性小鼠的基因治疗效果，我们将含有人β-Gal的AAV注射到小鼠脑内，并证明在一个月内仅在有限的区域检测到酶活性。我们将野生型人β-Gal转基因导入β-Gal KO小鼠，以挽救其GM 1-神经节苷脂沉积症表型。我们还将具有R201 C和I51 T氨基酸取代的突变型人β-Gal转基因引入β-Gal KO小鼠，这分别是幼年和成年GM 1-神经节苷脂沉积症常见的。使用了三种类型的含有β-肌动蛋白启动子/人β-Gal cDNA的转基因（野生型、R201 C或I51 T）。用β-Gal KO小鼠培育转基因小鼠，并且所得的三种类型的KO/转基因小鼠缺乏小鼠β-Gal基因，但具有人β-Gal（野生型，R201 C或I51 T）转基因。正如预期的那样，野生型人β-Gal转基因可以拯救小鼠GM 1-神经节苷脂沉积症。表达突变型人β-Gals的KO/转基因小鼠可能有助于研究具有残留酶活性的GM 1-神经节苷脂沉积症的发病机制和治疗。